Pharmaceutical composition for treating kidney cancer

Abstract

The invention relates to the technical field of biomedical engineering, and provides a pharmaceutical composition for treating kidney cancer, and the pharmaceutical composition comprises sunitinib, CYT387 and a pharmaceutically acceptable carrier. The invention also provides an application of the combination of sunitinib and CYT387 in preparation of a medicine for treating kidney cancer, and a result shows that the combination of CYT387 and sunitinib can obviously enhance the anti-kidney tumor activity and shows an obvious synergistic effect. Therefore, the invention provides a new exploration for overcoming the drug resistance of sunitinib, is beneficial to improving the drug sensitivity of malignant tumors, and has a certain clinical application prospect.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and more particularly to a drug for treating renal cancer, and more particularly to an application of a combination of sunitinib and CYT387 in treating renal cancer. Background Art

[0002] Renal cell carcinoma (RCC) is a highly malignant tumor in the urinary system and one of the most common tumors. It is a malignant tumor originating from the urinary tubular epithelial system of the renal parenchyma, also known as renal adenocarcinoma, accounting for 80% to 90% of renal malignant tumors. According to surveys, renal cancer ranks second among urogenital tumors in my country, second only to bladder tumors, accounting for 2% to 3% of adult malignant tumors and about 20% of pediatric malignant tumors. Renal cancer is classified into clear renal cell carcinoma, papillary renal cell carcinoma, chromophobe renal cell carcinoma and Bellini collecting duct carcinoma, accounting for about 90%. Clear renal cell carcinoma is the most common type, accounting for about 85%.

[0003] As a classic tyrosine kinase inhibitor, sunitinib is a multi-target anti-angiogenic drug that targets vascular endothelial growth factor (VEGF), thereby inducing apoptosis of renal cancer cells, inhibiting the growth of renal cancer cells, and prolonging the survival of patients with advanced renal cancer. It shows great promise in the treatment of advanced / metastatic renal malignancies. However, clinical data show that a large number of renal cancer patients show primary resistance to sunitinib or develop acquired resistance to sunitinib during treatment, which in turn affects its ultimate efficacy. Sunitinib combined with other adjuvant chemotherapy regimens can synergistically improve the efficacy of sunitinib, but it can also lead to increased adverse reactions and even patients who cannot tolerate treatment. Therefore, new drugs need to be developed to treat renal cancer.

[0004] CYT387 is a JAK inhibitor that can specifically inhibit the activity of nuclear factor kappa B kinase subunit epsilon (IKBKE). The molecular formula of compound CYT387 is: C 23 H 22 N 6 O 2 , CAS No.: 1056634-68-4, and its general structural formula is shown in formula (Ⅰ):

[0005]

[0006] The target of CYT387's anti-tumor effect is the kappaB kinase ε inhibitor IKBKE. It specifically inhibits the expression of IKBKE protein, producing a series of biological effects such as inhibiting tumor proliferation and inducing tumor cell apoptosis. Therefore, treating tumors by inhibiting IKBKE function by CYT387 has become one of the hot topics in the development of anti-tumor drugs in recent years. Whether the combination of sunitinib and CYT387 treatment can help enhance the inhibitory effect of sunitinib on renal cancer growth has not been reported yet. Summary of the invention

[0007] The purpose of the present invention is to provide a new sunitinib synergistic regimen, and further provide a pharmaceutical composition for treating renal cancer, in view of the fact that renal cancer patients in the prior art show primary resistance to sunitinib or develop acquired resistance to sunitinib during treatment, and that sunitinib combined with other adjuvant chemotherapy regimens will also lead to increased adverse reactions and even patients cannot tolerate the treatment.

[0008] The first aspect of the present invention provides a drug combination consisting of sunitinib and CYT387.

[0009] The second aspect of the present invention provides the use of the drug combination in the preparation of anti-tumor drugs. CYT387 inhibits the function of IKBKE to treat tumors; sunitinib, as a classic tyrosine kinase inhibitor, is a multi-target anti-angiogenic drug that targets vascular endothelial growth factor (VEGF) to induce cancer cell apoptosis and inhibit the growth of cancer cells. The combination of the two can be used to prepare tumors that develop by the above mechanism.

[0010] The third aspect of the present invention particularly provides the use of the drug combination in the preparation of a drug for treating renal cancer, preferably the concentration ratio between sunitinib and CYT387 is 1:1.

[0011] The results of cell experiments showed that in renal cancer cells 769-P and 786-O, the growth of renal cancer cells was significantly inhibited after the use of the drug combination of sunitinib and CYT387, compared with the use of sunitinib or CYT387 alone, **p<0.01, ***p<0.001; the results of animal model experiments showed that in the nude mouse tumor model of renal cancer, the weight of the tumor was significantly reduced after the use of the drug combination of sunitinib and CYT387, compared with the use of sunitinib or CYT387 alone, without increasing side effects.

[0012] In a fourth aspect, the present invention provides the use of CYT387 in preparing a sunitinib synergist. Preferably, the use is to enhance the inhibitory effect of sunitinib on the growth of renal cancer.

[0013] In a fifth aspect, the present invention provides a drug for treating renal cancer, comprising an active component and a pharmaceutically acceptable excipient, wherein the active component is the drug combination described above.

[0014] In terms of the form of preparation, the pharmaceutical composition is an oral preparation or an injection preparation. Preferably, sunitinib is in the form of an oral preparation, and CYT387 is in the form of an injection preparation.

[0015] Beneficial protection and effects of the present invention:

[0016] The present invention provides an application of sunitinib and CYT387 for the synergistic treatment of renal cancer. When the two are used in combination, the anti-kidney tumor activity can be significantly enhanced, showing a significant synergistic effect. Therefore, the present invention provides a new exploration for improving the drug resistance of sunitinib, helps to improve the drug sensitivity of malignant tumors, and has a certain clinical application prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The results show that CYT387 and sunitinib can inhibit renal cancer cell line 769-P in vitro.

[0018] Figure 2 The in vitro inhibitory effects of CYT387 and sunitinib on renal cancer cell line 786-O.

[0019] Figure 3 The in vivo inhibitory effects of CYT387 and sunitinib on renal cancer cell 786-O. DETAILED DESCRIPTION

[0020] The following examples and experimental examples are provided to further illustrate the present invention and should not be construed as limiting the present invention. The examples do not include detailed descriptions of conventional methods, which are well known to those skilled in the art and are described in many publications.

[0021] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the present invention, and the preferred implementation methods and materials described in the specific implementation methods are for demonstration purposes only.

[0022] 1. The drug combination of CYT387 and sunitinib significantly inhibits the activity of renal cancer cells in vitro

[0023] The CCK-8 (Cell Counting Kit-8) method was used to evaluate the inhibitory activity of 5 μM CYT387, 5 μM sunitinib, and the combination of the two drugs on renal cancer cells.

[0024] Human renal cancer cells 769-P and 786-O in the logarithmic growth phase (stored in this research center: Institute of Fudan University Cancer Hospital) were digested with trypsin to prepare a single cell suspension. The cells were counted using a hemocytometer and diluted to a cell concentration of 5×10 3 100 μL of culture medium was inoculated into a 96-well cell culture plate. Three more wells were set up with no cells but only the same volume of culture medium for blank control of instrument zeroing.

[0025] The cells were placed at 37°C with 5% CO 2 Incubate in the incubator for 24 hours, then remove the supernatant, add 100 μL of samples diluted with cell culture medium (5 μM sunitinib, 5 μM CYT387, 5 μM sunitinib and 5 μM CYT387), add the same dilution of dimethyl sulfoxide (DMSO) and 100 μL culture medium to the control well and blank control well, respectively. After culturing for 2-4 days, replace the same volume of fresh cell culture medium containing 10% CCK-8 reagent. Incubate at 37°C for 2 hours and measure by colorimetric analysis with an enzyme reader (measurement wavelength 450nm).

[0026] The relative cell growth rate was calculated as follows:

[0027]

[0028] Test results: In renal cancer cells 769-P and 786-O, the combination of sunitinib and CYT387 significantly inhibited the growth of renal cancer cells compared with sunitinib or CYT387 alone, **p<0.01, ***p<0.001. For specific data, see Figure 1 , Figure 2 and Table 1:

[0029] Table 1 In vitro experiments on the inhibitory effect on prostate cancer cells

[0030]

[0031] 2. Nude Mouse Tumor Experiment

[0032] 1. Construction of mouse model

[0033] 786-O cells (stored in our research center: Institute of Cancer Hospital Affiliated to Fudan University) were used to subcutaneously implant nude mice (BALB / cNude, purchased from Beijing Sibeifu) until the tumor volume reached 50mm 3 After that, the preparation is completed.

[0034] 2. Formula for injection for treating kidney cancer

[0035] Sunitinib was prepared into an oral preparation, and CYT387 was prepared into an injection. The specific formulas are shown in Table 2:

[0036] Table 2 Formula of injection for treating renal cancer

[0037]

[0038] Preparation method:

[0039] Sunitinib was dissolved in dimethyl sulfoxide (DMSO), and then polyethylene glycol 300, Tween-80, and water for injection were added in sequence, and then ultrasonicated to prepare a uniform suspension with an active ingredient content of 1 mg / mL;

[0040] CYT387 was dissolved in dimethyl sulfoxide (DMSO), and then polyethylene glycol 300, Tween-80, and water for injection were added in sequence and mixed into a clear liquid with an active ingredient content of 1 mg / mL.

[0041] 3. Tumor volume test

[0042] When the tumor volume of mice reaches 50mm 3 Afterwards, sunitinib (0.2 mg / 10 g / 2 d, oral, oral agent) and / or CYT387 (1 mg / 10 g / 2 d, intraperitoneal injection, injection) were administered, and the blood vessels were taken out for measurement after 4 weeks.

[0043] Test results: In the nude mouse model of renal cancer, the volume and weight of the tumor were significantly reduced after the drug combination of sunitinib and CYT387 was applied, ***p<0.001. For specific experimental data, see Figure 3 and Table 3.

[0044] Table 3 Inhibitory effect of in vivo experiments on prostate cancer cells

[0045] Group Tumor weight (g) Weight of nude mice (g) Comparison 1.05±0.25 21.93±0.91 Sunitinib 0.62±0.17 21.43±0.88 CYT487 0.51±0.13 21.88±1.25 Sunitinib + CYT487 0.21±0.08 21.53±1.29

[0046] The results show that in the nude mouse model of renal cancer, the weight of the tumor was significantly reduced after the combination of sunitinib and CYT387 was used compared with the use of sunitinib or CYT387 alone, and the mice did not experience serious adverse reactions and their body weight did not change significantly. This shows that the combination of sunitinib and CYT387 can enhance the therapeutic effect without increasing side effects.

[0047] The preferred embodiments of the present invention have been specifically described above, but the present invention is not limited to the embodiments. Those skilled in the art may make various equivalent modifications or substitutions without violating the spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. A drug combination, characterized in that Composed of sunitinib and CYT387.

2. The drug combination according to claim 1, characterized in that The concentration ratio between sunitinib and CYT387 was 1:

1.

3. Use of the combination of sunitinib and CYT387 in the preparation of anti-tumor drugs.

4. Use of the combination of sunitinib and CYT387 in the preparation of drugs for the treatment of renal cancer.

5. A pharmaceutical composition for treating renal cancer, characterized in that: The invention comprises an active component and a pharmaceutically acceptable excipient, wherein the active component is the drug combination according to claim 1 or 2.

6. The pharmaceutical composition for treating renal cancer according to claim 5, characterized in that: The pharmaceutical composition is an oral preparation or an injection preparation.

7. Application of CYT387 in the preparation of sunitinib synergist.

8. The use according to claim 7, characterized in that: The application is to improve the inhibitory effect of sunitinib on the growth of renal cancer.

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